Unique Plant Material for Studying Woody-plant Nitrogen Metabolism

Temperate woody perennials produce proteins in the stem for seasonal nitrogen (N) storage. In Populus species, this seasonal N storage occurs primarily as a 32-kDa Bark Storage Protein (BSP), which can accumulate to 50% of total bark proteins during the winter. Plants of the Populus tremula × Populus alba (clone 717) were transformed with the BSP cDNA in antisense orientation (fused to a constitutive promoter), and regenerated lines were screened. Several independent antisense-BSP (A-BSP) lines were selected, which, after 4 weeks of SD photoperiod, showed 70% to 90% reduction in total BSP accumulation compared to the wild-type (WT). A series of experiments were conducted to compare LD growth of one A-BSP line to that of the WT. A-BSP plants showed reduced growth at both 5 and 50 mM ammonium nitrate fertilization. However, the higher N rate eventually resulted in toxicity in WT, but not in A-BSP plants. A-BSP plants grown hydroponically (0.5x Hoagland1s) showed altered partitioning with reduced stem length and increased leaf area (Leaf:stem dry-weight ratios were 14.8 and 20.9 for ABSP and WT, respectively). Partitioning to the roots was not different between A-BSP and WT. Proposed functions of BSP in seasonal and LD nitrogen metabolism will be discussed.

Bark storage Protein in Peach, Plum, and Cherry Trees

The main form of nitrogen reserves during overwintering are amino acids and proteins. Specific proteins called bark storage proteins (BSP) have been characterized in many tree species. To identify BSPs in `O'Henry' peach, `Angeleno' plum, and `Early Burlat' cherry trees, samples of bark were collected from January through December 1993 from trees growing under field conditions in Santiago, Chile. SDS-PAGE analyses were used to characterize the seasonal variation on the protein pattern on the bark of those Prunus species. A 60 kDa BSP was identified in the bark of all three species, which corresponds to the main protein present in the bark during the winter. This protein may play an important role as a nitrogen reserve in these fruit trees.

COLD ACCLIMATION IN THE LEAVES OF SIBLING DECIDUOUS AND EVERGREEN PEACH: ALTERATIONS IN DEHYDRIN AND BARK STORAGE PROTEINS

Seasonal pattern of cold tolerance and proteins were studied in the leaves of sibling deciduous and evergreen peach (Prunus persica). In contrast to deciduous peach that undergoes endodormancy in fall, evergreen peach does not (leaves are retained and shoot tips elongate under favorable conditions) (Arora et al., Plant Physiol. 99:1562-1568). Cold tolerance (LT50) was assessed using electrolyte leakage method. Proteins were separated by SDS-PAGE. Electroblots were probed with anti-dehydrin (Dr. T. Close) and anti-19 kD, peach bark storage protein (BSP) antibodies. LT50 of leaves successively increased from about -7C (18 Aug.) to -15C and -11.5C (23 Oct.) in deciduous and evergreen genotypes, respectively. The most apparent change in the protein profiles was the accumulation of a 60-kD protein during cold acclimation in the leaves of deciduous trees; however, it did not change significantly in evergreen peach. Immunoblots indicate that 60-kD protein is a dehydrin protein. PAGE and immunoblots indicated that 19-kD BSP disappeared progressively during summer through fall in the leaves of deciduous peach, but accumulated to large amounts in bark tissues. Similar inverse relationship for its accumulation in leaf vs. bark tissue was not evident in evergreen peach. Results indicate that BSP expression may be regulated by altered source/sink relationship.

DEGRADATION OF A BARK STORAGE PROTEIN IN POPLAR.

A 32kDa bark storage protein (BSP) which accumulates in the fall and is degraded in the spring has been identified in Populus deltoides bark. The BSP gene has been shown to be regulated by short day (SD) photoperiod (8 h). The physiological condition of the plant and the environmental factors necessary for the degradation and retranslocation of BSP are of considerable interest for determining the role of this protein in the remobilization of nitrogen in trees. Poplar plants were placed in a SD growth chamber for 4 or 7 weeks to induce growth cessation (bud set) or dormancy, respectively. BSP accumulated to high levels in bark tissues after 3 weeks SD and remained high through 7 weeks SD. Plants in which growth had stopped (4 weeks SD), or in which dormancy (7 weeks SD) was broken with hydrogen cyanamide (0.5 M) or chilling (4 weeks 0C) broke bud within 1 week of being placed into long day (LD) conditions. Dormant plants which were not chilled broke bud after 3 weeks LD. BSP levels decreased around the time of budbreak, suggesting that the degradation of BSP is dependent on the need for a nitrogen sink, ie. budbreak and new shoot growth.